The oxygen radicals involved in the toxicity induced by parthenolide in MDA-MB-231 cells
The oxygen radicals involved in the toxicity induced by parthenolide in MDA-MB-231 cells
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DOI：
10.3892/or.2014.3212
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发表时间：
2014-07-01
期刊：
影响因子：
4.2
通讯作者：
Lauricella, Marianna
中科院分区：
文献类型：
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作者：
Carlisi, Daniela;D'Anneo, Antonella;Lauricella, Marianna
研究表明，倍半内酯内酯降低了MDA-MB-231乳腺癌细胞的活力，这与氧化应激有关。本报告考察了在巴特内酯治疗过程中产生的不同自由基物种，以及它们在药物引起的毒性中可能发挥的作用。时程实验表明，在处理的第一阶段(0-8h)，特别是在前3h，菊内酯在很大比例的细胞中诱导了二氯荧光素(DCF)信号，而二氢乙锭(DHE)信号不被激发。由于对DCF信号的影响被NADPH氧化酶(NOX)的两种阻断剂apocynin和DPI所抑制，我们认为，Parthenolide通过产生超氧阴离子(O-2(中心点-))迅速刺激NOx活性，而超氧化物歧化酶1(SOD 1)将其转化为过氧化氢(H_2O_2)。在第二阶段(8-16h)，菊内酯增加了对DHE信号的阳性细胞数。由于这一事件不能被apocynin和DPI阻止，并且与细胞对MitoSox Red的阳性有关，MitoSox Red是一种用于检测线粒体产生O-2(中心点-)的荧光色素，我们认为在治疗的第二阶段，Parthenolide独立地通过NOX活性在线粒体水平诱导O-2(中心点-)的产生。最后，在这个阶段，大多数细胞对羟基苯基荧光素(HPF)信号呈阳性，这是一种检测高活性氧物种(HROS)的荧光探针，如羟基自由基和过氧亚硝酸根。因此，在处理8-16h之间，菊内酯诱导产生O-2(中心点-)和hros，这与细胞存活率的显著下降密切相关。
It has been shown that the sesquiterpene lactone parthenolide lowers the viability of MDA-MB-231 breast cancer cells, in correlation with oxidative stress. The present report examined the different radical species produced during parthenolide treatment and their possible role in the toxicity caused by the drug. Time course experiments showed that in the first phase of treatment (0-8 h), and in particular in the first 3 h, parthenolide induced dichlorofluorescein (DCF) signal in a large percentage of cells, while dihydroethidium (DHE) signal was not stimulated. Since the effect on DCF signal was suppressed by apocynin and diphenyleneiodonium (DPI), two inhibitors of NADPH oxidase (NOX), we suggest that parthenolide rapidly stimulated NOX activity with production of superoxide anion (O-2(center dot-)), which was converted by superoxide dismutase 1 (SOD1) into hydrogen peroxide (H2O2). In the second phase of treatment (8-16 h), parthenolide increased the number of positive cells to DHE signal. Since this event was not prevented by apocynin and DPI and was associated with positivity of cells to MitoSox Red, a fluorochrome used to detect mitochondrial production of O-2(center dot-), we suggest that parthenolide induced production of O-2(center dot-) at the mitochondrial level independently by NOX activity in the second phase of treatment. Finally, in this phase, most cells became positive to hydroxyphenyl fluorescein (HPF) signal, a fluorescent probe to detect highly reactive oxygen species (hROS), such as hydroxyl radical and peroxynitrite. Therefore, parthenolide between 8-16 h of treatment induced generation of O-2(center dot-) and hROS, in close correlation with a marked reduction in cell viability.